1.Identification of the Antioxidant of Chrysanthemum morifolium Using Entropy-weighted TOPSIS Method Combined with Spectrum-effect Analysis
Qianqian ZHU ; Shiqin WANG ; Xuxiang ZHOU ; Ruili NIE ; Jianting ZHAN ; Bo WANG ; Jing NIE ; Xiaochuan YE ; Dan LIU
Herald of Medicine 2024;43(6):956-964
Objective To evaluate the antioxidant function of Chrysanthemum morifolium from different origins and to identify their antioxidant material basis.Methods The HPLC fingerprints of the water extracts of C.morifolium from different origins were established.The antioxidant activities of C.morifolium were assayed by measuring the 2.2-diphenyl-l-picrylhydrazyl(DPPH),hydroxyl radical,ABTS,superoxide anion radical scavenging capacity and ferric ion reducing capacity FRAP.Entropy-weighted TOPSIS was used to calculate the weighting coefficients of the single indexes.Grey relational analysis(GRA)and partial least squares were used for spectrum-effect analysis to identify the antioxidant material basis of C.morifolium.Results A total of 16 common peaks were discovered in the fingerprint of the water extracts of 10 batches of C.morifolium,among which 13 common components were identified.All the C.morifolium samples had good antioxidant capacity,and the results of entropy-weighted TOPSIS analysis showed that the ranking of total antioxidant potency of 10 batches of C.morifolium was follows:S1>S8>S3>S5>S4>S10>S7>S2>S9>S6.The peaks of 1-5,9,10,12,14 were positively correlated with the antioxidant activity and the variable influence on projection(VIP)values were greater than 1.The correlation coefficients of these nine peaks in GRA were all greater than 0.7.Conclusion The entropy-weighted TOPSIS method combined with the spectrum-effect analysis could be used to screen out the antioxidant material basis of C.morifolium and the results provide a basis for establishing quality assessment system for C.morifolium based on Quality-markers thus improving the quality control level.
2.Screening of Anti-Helicobacter pylori Gastritis Active Component of the Ethyl Acetate Extract of Alpinia officinarum Hance Based on the Knockout Strategy
Xin LUO ; Wuyingxiao ZHENG ; Jianting ZHAN ; Jingyu YANG ; Haoran MA ; Guoping GAN ; Pengtao YOU ; Xiaochuan YE ; Dan LIU
Herald of Medicine 2024;43(9):1387-1392
Objective To screen the anti-Helicobacter pylori gastritis active components of the ethyl acetate extract of Alpinia officinarum Hance.Methods The"knock-out"strategy combined with high-performance liquid chromatography(HPLC)detection was developed to separate the components of the ethyl acetate extract of A.officinarum while obtaining the negative samples without the components.A human gastric epithelial cell(GES-1)model of H.pylori gastritis was established,and the levels of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),interleukin-8(IL-8)and interleukin-1β(IL-1β)in the supernatant of the cells were determined by enzyme-linked immunosorbent assay(ELISA).Results The total flavonoid fraction,the negative fraction without total diphenylheptanoids,the negative fraction without 5-hydroxy-7-(4-hydroxy-3-methoxyphenyl)-1-phenyl-3-heptanone(DHPA),and galangin significantly reduced IL-6 levels in the supernatant of H.pylori infected GES-1 cells at a concentration of 8 μg·mL-1 with 24 h incubation.The total flavonoid fraction strongly inhibited the release of IL-6,TNF-α,IL-8,and IL-1β from H.pylori gastritis GES-1 cells at a concentration of 16 μg·mL-1.Conclusions The total flavonoid fraction is the major anti-H.pylori gastritis active component of the ethyl acetate extract of A.officinarum.The results lay the foundation for further elucidation of the material basis of A.officnarum against H.pylori gastritis.
3.Protective Effect against Helicobacter pylor Gastritis in Mice by Flavonoid Combinations of Alpiniae Officinarum Rhizoma via Inhibition of PI3K/Akt Pathway
Xin LUO ; Wuyinxiao ZHENG ; Jingyu YANG ; Jianting ZHAN ; Haoran MA ; Xiaochuan YE ; Guopin GAN ; Dan LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(8):61-68
ObjectiveTo investigate the protective effect and mechanism of action of flavonoid combination of Alpiniae Officinarum Rhizoma (A. officinarum) against Helicobacter pylori (H. pylori) gastritis in mice. MethodsAfter acclimatization for one week, 56 SPF-grade healthy C57BL/6J mice were gavaged with mixed antibiotics for three consecutive days. They were randomly divided into a normal group, model group, positive drug group (triple therapy group), and low- and high-dose groups (100, 200 mg·kg-1) of flavonoid combination of A. officinarum. The H. pylori gastritis mice model was established by gavage with H. pylori bacterial suspension in each group except for the normal group. After successful modeling, mice were administrated with corresponding drugs once a day for two weeks. Hematoxylin-eosin (HE) staining was used to observe the pathological changes in gastric tissue. Rapid urease test paper was used to detect the positive rate of H. pylori. Silver staining was used to observe the H. pylori adherence on the surface of gastric tissue. Immunohistochemistry was used to detect the protein expression of interleukin-8 (IL)-8 and myeloid differentiation factor (MyD88) in gastric tissue. The serum levels of IL-6, tumor necrosis factor-α (TNF-α), IL-8, and IL-1β were detected by enzyme-linked immunosorbent assay (ELISA). The expressions of phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) protein were detected by Western blot. ResultsCompared with those in the normal group, mice in the model group had lower gastric weight coefficients, higher pH of gastric juice, 100% H. pylori infection rate, and significantly changed gastric histopathology. The expressions of IL-8 and MyD88 proteins in the gastric tissue of mice in the model group were significantly elevated, and the serum levels of inflammatory factors IL-6, TNF-α, IL-8, and IL-1β were significantly up-regulated in mice. Compared with that in the model group, the gastric weight coefficient of mice in each treatment group of the flavonoid combinations of A. officinarum was elevated (P<0.01), and the pH of gastric juice was reduced (P<0.01). The infection rate of H. pylori was reduced. The expressions of IL-8 and MyD88 proteins in the gastric tissue of mice in the treatment groups were significantly reduced (P<0.01), and the serum levels of inflammatory factors IL-6, TNF-α, IL-8, and IL-1β were significantly reduced in a dose-dependent manner (P<0.01). The flavonoid combinations of A. officinarum down-regulated the expression of PI3K and Akt proteins in H. pylori gastritis-infected cells (P<0.01). ConclusionThe protective effect of flavonoid combinations of A. officinarum against H. pylori gastritis is associated with the inhibition of H. pylori infection rate and regulation of PI3K/Akt signaling pathway, resulting in inhibiting the release of inflammatory factors.